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《Chinese Journal of Clinical Rehabilitation》 2006-43
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Interventional effect of resibufogenin on the cell growth of human gastric cancer BGC-823 cell line

Meng Shu-cong1, Dong Xiao-min1, Xiao Jun-jun1, Wang Chuan-she2, Guo De-an2 1Department of Cell Biology, Basic Medical College, Peking University, Beijing 100083, China; 2Chinese Pharmaceutical Science Research Center, Peking University Medical Science Center, Beijing 100083, China  
AIM: To observe the intervention of resibufogenin, one of main components of Chinese herb venenum bufonis, on the growth of human gastric cancer BGC-823 cell line cell cultured in vitro, and the relationship with cytochrome C and Caspase-3 activation. METHODS: The experiment was conducted in the Laboratory of Department of Cell Biology, Basic Medical College, Peking University from September 2004 to December 2005. BGC-823 cell line was cultured in vitro. Resibufogenin of different concentrations were added in each resibufogenin group (Firstly, dissolved with dimethyl sulphoxide, and then diluted with RPMI-1640 culture solution till final concentration; resibufogenin was 0.1, 1, 10 μmol/L in the cell proliferation and proliferation inhibition test, morphous observation of cell nucleus and DNA content determination tests; resibufogenin was 0, 1, 2.5, 5 μmol/L in the cell-cycle distribution test, cell apoptosis test, mitochondria membrane potential test). RPMI-1640 culture solution was added in the blank control group. 5 μmol/L aclarubicin hydrochloride was added in the aclarubicin hydrochloride group. Anti-proliferation effect was measured with acid phosphatase assay (APA). Cell morphological change and DNA content determination were observed with cell fluorescent spectrophotometer. Cell cycle, apoptotic rate and changes in mitochondrial transmembrane potential were measured with cytofluorometry. Change of cell apoptosis related gene protein was determined with Western blot. RESULTS: ①After the BGC-823 cells were treated with 0.1, 1, 10 μmol/L resibufogenin for 24, 48 and 72 hours, the cell growth was inhibited significantly. The percentage of resibufogenin on grow inhibiting of cancer cell had positive correlation with dosage and time, and IC50 was 3.9, 2.0, 1.5 μmol/L, respectively. ②With the prolongation of time and increase of concentration of resibufogenin, fluorescence intensity of cancer cell nucleus reduced and the content of cell nucleus DNA decreased obviously. ③0.1, 1 μmol/L resibufogenin induced accumulation of cells in S phase after being cultured for 24-48 hours. The 5 μmol/L resibufogenin induced accumulation of cells in G2+M phase after being cultured for 72 hours. Interaction between aclarubicin hydrochloride and cancer cell induced accumulation of cells in S phase 24-48 hours later, in G2+M phase for 72 hours. ④After resibufogenin effect, the apoptotic rate of cells was markedly higher than that in the control group. ⑤Resibufogenin could induce the decrease of cell mitochondria membrane potential, increase cytochrome C in kytoplasm, enhance Caspase 3 activation, inhibit the expression of Bcl-2 protein and result in cell apoptosis. CONCLUSION: Resibufogenin can inhibit the growth of BGC-823 cell and induce the apoptosis of BGC-823 cells in vitro. The mechanism of inducing apoptosis is associated with the mitochondrial dependent pathway.
【Fund】: 国家自然科学基金(330690);; 教育部教育振兴行动计划专项(985-2-016-24)~~
【CateGory Index】: R735.2
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